IS1071-mediated recombinational equilibrium in Alcaligenes sp. BR60 carrying the 3-chlorobenzoate catabolic transposon Tn5271

Abstract

In experiments designed to Tn5 mutagenize the indigenous plasmid pBRC60 of Alcaligenes sp. BR60, kanamycin-resistant mutants were isolated that were cured of this plasmid and that exhibited recombination of the plasmid-located chlorobenzoate catabolic transposon Tn5271 into the chromosome. These events were independent of the location of Tn5 insertions into the genome of strain BR60. The chromosomal recombinants carried at least two novel copies of IS1071, the class II insertion sequence flanking Tn5271, compared with the parent strain. Recombination of Tn5271 into the chromosome of Alcaligenes sp. BR60 was also detected following mating in of pBRC60-incompatible (IncP1) plasmids, R68 and pGS65. Chromosomal copies of Tn5271 could be mobilized between Alcaligenes strains via plasmids pBRC40 or R68. Conjugation of the incompatible plasmid pGS65 into Alcaligenes strains in the absence of selection for 3-chlorobenzoate catabolism resulted in the recovery of 85% of transconjugants in which the entire pBRC60 plasmid had integrated into the chromosome. These transconjugants exhibited complex rearrangements in chromosomal IS1071 copies. A model of recombinational equilibrium involving homologous recombination between plasmid and chromosomal copies of IS1071 is presented. The results are discussed in terms of the IS1071 (class II) transposition mechanism and the observed products of IS 1071-mediated recombination in natural recipients of pBRC60 in aquatic environments.Key words: transposon, 3-chlorobenzoate catabolism, rearrangement.